Extracellular vesicles(EVs)are vesicles with phospholipid bilayer structure actively secreted by cells,which naturally carry various contents derived from mother cells.EVs retain the biological char-acteristics of mother cells and reflect the status of mother cells during their secretion.In order to under-stand the biogenesis of EVs and their biological functions in the body,scientists have made many at-tempts.However,direct detection of EVs has some difficulties and current detection techniques usually use EVs contents as detection objects to achieve the purpose of indirect characterization.The purpose of disease diagnosis can be achieved by analyzing EVs in body fluids,and detailed characterization of EVs can promote the development of precision medicine.EVs belong to endogenous substances and can be used as carriers to deliver drugs,with the advantages of increasing efficacy,reducing drug toxicological effects and passing through biological barriers.When EVs applied as a drug or a carrier for delivering drug,challenges to be addressed include(ⅰ)clear characterization of the contents of EVs,(ⅱ)develop-ment of suitable assays to monitor EVs in vivo to determine and optimize the dose,route of administra-tion,and potential toxicity of EVs,and (ⅲ) determination of the kind and amount of membrane compo-sition of EVs.The diameter of EVs is nanoscale and the content is small,but it is of great value in clini-cal application.The selection of appropriate analytical techniques in the application will enable a compre-hensive characterization of EVs and further advance our understanding of EVs biology to develop novel di-agnostic and therapeutic approaches.In this review,the principles,applications,advantages and disad-vantages of analytical techniques in the study of EVs are introduced,with emphasis on the characteriza-tion techniques of EVs.

Objective:To evaluate the effect of electroacupuncture on liver regeneration after partial hepatectomy in mice and the role of the Notch signaling pathway.Methods:Thirty-six SPF healthy male C57BL/6 mice, aged 6 weeks, weighing 20-22 g, were divided into 6 groups ( n=6 each) using a random number table method: sham operation group (group S), partial hepatectomy group (group PH), non-acupoint electroacupuncture+ partial hepatectomy group (group NPH), partial hepatectomy+ Fli-06 group (group PH+ F), acupoint electroacupuncture+ partial hepatectomy group (group EPH), and acupoint electroacupuncture+ partial hepatectomy+ Fli-06 group (group EPH+ F). All the mice except for group S underwent partial hepatectomy. Fli-06 4.8 mg/kg was intraperitoneally injected starting from 2 days before surgery, once a day, until the mice were sacrificed in group PH+ F and group EPH+ F, while the equal volume of 0.9% sodium chloride solution was injected in the other groups. In EPH group, electroacupuncture of bilateral " Zusanli" acupoints lasting for 15 min was performed using continuous waves with a frequency of 2 Hz and an intensity of 1 mA once a day starting from the time point immediately after surgery for 3 consecutive days. Mice were anesthetized at day 2 after partial hepatectomy, and blood samples were taken from the eyeball for determination of the serum alanine transaminase (ALT) and aspartate aminotransferase (AST) concentrations (using a fully automated biochemical analyzer) and concentrations of serum epidermal growth factor (EGF) and hepatocyte growth factor (HGF) (by enzyme-linked immunosorbent assay). The mice were subsequently sacrificed and liver tissues were taken for calculation of the liver mass to body mass ratio and for determination of the expression of liver proliferation marker Ki-67 (by immunohistochemical staining), proliferating cell nuclear antigen (PCNA), cyclin D1 (CCND1), Notch Intracellular Domain (NICD), and hypoxia-inducible factor-1alpha (HIF-1α) (using Western blot) and Notch1, jagged canonical Notch ligand 1 (Jagged1) and hairy and enhancer of split 1 (Hes1) mRNA (by real-time polymerase chain reaction). Results:Compared with group S, the serum ALT, AST, EGF and HGF concentrations were significantly increased, and the expression of hepatic Notch1, Jagged1 and Hes1 mRNA and Ki-67, PCNA, CCND1 and NICD was up-regulated in group PH ( P<0.05 or 0.01). Compared with group PH, the liver mass to body mass ratio and serum EGF and HGF concentrations were significantly increased, the serum ALT and AST concentrations were decreased, and the expression of hepatic Notch1, Jagged1, Hes1 mRNA and Ki-67, PCNA, CCND1, NICD and HIF-1α was up-regulated in group EPH, and the liver mass to body mass ratio and the serum HGF concentrations were significantly decreased, the serum ALT and AST concentrations were increased, and the expression of hepatic Jagged1 and Hes1 mRNA and Ki-67, PCNA, CCND1, NICD, and HIF-1α was down-regulated in group PH+ F ( P<0.05 or 0.01). Compared with group EPH, the liver mass to body mass ratio and serum EGF and HGF concentrations were significantly decreased, the serum ALT and AST concentrations were increased, and the expression of hepatic Notch1, Jagged1, Hes1 mRNA and Ki-67, PCNA, CCND1, NICD and HIF-1α was down-regulated in group EPH+ F ( P<0.01). Conclusions:Electroacupuncture at Zusanli acupoint promotes liver regeneration after partial hepatectomy in mice, and the mechanism may be related to the activation of the Notch signaling pathway.

AIM To study the Chemical constituents from Rhinacanthus nasutus(L.)Kurz and their in vitro antitumor and lipid-lowering activities.METHODS The ethyl acetate fraction from R.nasutus was isolated and purified by normal phase,reverse phase silica gel column and semi-preparative HPLC,then the structures of obtained compounds were identified by physicochemical properties and spectral data.The in vitro anti-tumor activity was determined by MTT assay,and the in vitro lipid-lowering activity was evaluated by oleic acid-induced HepG2 high-fat cell model.RESULTS Twenty compounds were isolated and identified as rhinacanthin A(1),rhinacanthin B(2),rhinacanthin C(3),rhinacanthin D(4),rhinacanthin E(5),rhinacanthin M(6),rhinacanthin N(7),rhinacanthin Q(8),rhinacanthin T(9),rhinacanthone(10),β-sitosterol(11),gallic acid(12),vanillic acid(13),syringic acid(14),lapachol(15),umbelliferone(16),sambucunlin A(17),17α,21-dihydroxy-1,4-pregnadiene-3,11,20-trione,21-acetate(18),1,8-dihydroxyanthraquinone(19),2,6-dimethoxy-1,4-benzoquinone(20).Compounds 3 and 7 inhibited the proliferation of HepG2,Hela,A549 and H22 tumor cells with IC50 values of(0.66±0.17)-(3.22±0.49)μmol/L in a time-and concentration-dependent manner.Compounds 3,7 and 9 had a lipid clearance rate of more than 50%in oleic acid-induced HepG2 high-fat cells.CONCLUSION Compounds 16-20 are isolated from this plant for the first time.Compounds 3 and 7 have good in vitro anti-tumor activities,and 3,7,9 have good in vitro lipid-lowering activities.

Objective To investigate the incidence rate,clinical characteristics,and prognosis of neonatal stroke in Shenzhen,China.Methods Led by Shenzhen Children's Hospital,the Shenzhen Neonatal Data Collaboration Network organized 21 institutions to collect 36 cases of neonatal stroke from January 2020 to December 2022.The incidence,clinical characteristics,treatment,and prognosis of neonatal stroke in Shenzhen were analyzed.Results The incidence rate of neonatal stroke in 21 hospitals from 2020 to 2022 was 1/15 137,1/6 060,and 1/7 704,respectively.Ischemic stroke accounted for 75％(27/36);boys accounted for 64％(23/36).Among the 36 neonates,31(86％)had disease onset within 3 days after birth,and 19(53％)had convulsion as the initial presentation.Cerebral MRI showed that 22 neonates(61％)had left cerebral infarction and 13(36％)had basal ganglia infarction.Magnetic resonance angiography was performed for 12 neonates,among whom 9(75％)had involvement of the middle cerebral artery.Electroencephalography was performed for 29 neonates,with sharp waves in 21 neonates(72％)and seizures in 10 neonates(34％).Symptomatic/supportive treatment varied across different hospitals.Neonatal Behavioral Neurological Assessment was performed for 12 neonates(33％,12/36),with a mean score of(32±4)points.The prognosis of 27 neonates was followed up to around 12 months of age,with 44％(12/27)of the neonates having a good prognosis.Conclusions Ischemic stroke is the main type of neonatal stroke,often with convulsions as the initial presentation,involvement of the middle cerebral artery,sharp waves on electroencephalography,and a relatively low neurodevelopment score.Symptomatic/supportive treatment is the main treatment method,and some neonates tend to have a poor prognosis.

Aim To explore the effects of Lycium berry seed oil on Nrf2/ARE pathway and oxidative damage in testis of subacute aging rats. Methods Fifty out of 60 male SD rats, aged 8 weeks, were subcutaneously injected with 125 mg • kg"D-galactosidase in the neck for 8 weeks to establish a subacute senescent rat model. The presence of senescent cells was observed using P-galactosidase ((3-gal), while testicular morphology was examined using HE staining. Serum levels of testosterone (testosterone, T), follicle-stimulating hormone ( follicle stimulating hormone, FSH ) , luteinizing hormone ( luteinizing hormone, LH ) , superoxide dis-mutase ( superoxide dismutase, SOD ) , glutathione ( glutathione, GSH) and malondialdehyde ( malondial-dehyde, MDA) were measured through ELISA, and the expressions of factors related to aging, oxidative damage, and the Nrf2/ARE pathway were assessed via immunohistochemical analysis and Western blotting. Results After successfully identifying the model, the morphology of the testis was improved and the intervention of Lycium seed oil led to a down-regulation in the expression of [3-gal and -yH2AX. The serum levels of SOD, GSH, T, and FSH increased while MDA and LH decreased (P 0. 05) . Additionally, there was an up-regulated expression of Nrf2, GCLC, NQOl, and SOD2 proteins in testicular tissue ( P 0. 05 ) and nuclear expression of Nrf2 in sertoli cells. Conclusion Lycium barbarum seed oil may reduce oxidative damage in testes of subacute senescent rats by activating the Nrf2/ARE signaling pathway.

Chronic obstructive pulmonary disease (COPD) is the most common chronic airway disease. The current status of treatment based mainly on bronchodilators and ICS is not sufficient for all of COPD patients. Various studies have attempted to use biologics targeting specific cytokines and their receptors in COPD patients to alleviate respiratory symptoms or reduce the risk of acute exacerbations. However, they failed to bring significant clinical benefits. More studies are needed to further determine the efficacy of targeted biotherapy for COPD.

Chronic cough is a common condition that imposes significant physical, psychological, and social burdens on patients. Although chronic cough is often associated with underlying conditions such as asthma, gastroesophageal reflux disease, and eosinophilic bronchitis, some patients experience uncontrollable coughing that is difficult to attribute to a specific cause. Many of these patients exhibit clinical features of cough hypersensitivity syndrome, providing new directions for research into the treatment of chronic cough. As the pathophysiological mechanisms of chronic cough are further elucidated, treatment approaches for chronic cough are entering a new stage of development. This article summarizes and discusses the mechanisms and clinical evidence of central neuromodulators used in the treatment of chronic cough, suggesting promising clinical applications for these drugs in the future.

Objective To explore the characteristics of blood lipid metabolism indicators and risk factors of prognosis in children with systemic lupus erythematosus (SLE). Methods A total of 54 children who were diagnosed with SLE and hospitalized in Chengdu Women and Children’ s Central Hospital from January 2013 to August 2022 were selected. Clinical data of all children were collected and blood lipid metabolism indicators and biochemical indicators were detected , and binary logistic regression was used to analyze the prognosis risk factors in children with SLE. Results Among the 47 cases (87.04%) had abnormal blood lipid metabolism at admission, and is mainly manifested as elevated levels of LDL-C, TG and TC and decreased level of HDL-C. The proportion of cardiovascular system damage, hematological system damage, urinary protein positivity, and SLEDAI-2000 score in the group with good prognosis were lower than those in the group with poor prognosis, while the proportion of dsDNA positivity was higher in the group with poor prognosis. Binary Logistic regression analysis showed that the cardiovascular system damage and positive urinary protein were risk factors for poor prognosis, with statistically significant differences (P<0.05). Conclusion Abnormal blood lipid metabolism is common in children with SLE, and cardiovascular system damage and positive urinary protein may increase the risk of poor prognosis in young children.

Chronic obstructive pulmonary dis-ease(COPD)is the most common chronic airway disease.The current status of treatment based mainly on bronchodilators and ICS is not sufficient for all of COPD patients.Various studies have at-tempted to use biologics targeting specific cyto-kines and their receptors in COPD patients to allevi-ate respiratory symptoms or reduce the risk of acute exacerbations.However,they failed to bring significant clinical benefits.More studies are need-ed to further determine the efficacy of targeted biotherapy for COPD.

Chronic cough is a common condition that imposes significant physical,psychological,and social burdens on patients.Although chronic cough is often associated with underlying condi-tions such as asthma,gastroesophageal reflux dis-ease,and eosinophilic bronchitis,some patients ex-perience uncontrollable coughing that is difficult to attribute to a specific cause.Many of these pa-tients exhibit clinical features of cough hypersensi-tivity syndrome,providing new directions for re-search into the treatment of chronic cough.As the pathophysiological mechanisms of chronic cough are further elucidated,treatment approaches for chronic cough are entering a new stage of develop-ment.This article summarizes and discusses the mechanisms and clinical evidence of central neuro-modulators used in the treatment of chronic cough,suggesting promising clinical applications for these drugs in the future.